An immunogenetic analysis of T-cell reactive regions on the major allergen from the house dust mite, Der p I, with recombinant truncated fragments.

Abstract

Proliferation assays were used to localize the T-cell reactive sites on the major allergen from the house dust mite Dermatophagoides pteronyssinus, Der p I. Seven overlapping recombinant fragments of the Der p I molecule, synthesized with the pGEX expression vector system, were used to stimulate peripheral blood lymphocytes from 35 HDM-sensitive individuals. The fusion fragments were from 39 to 114 amino acids in length and spanned the entire Der p I molecule. Significant proliferative responses to one or more of the fragments were evident in 18 of the allergic individuals, and each of the fragments led to T-cell stimulation in at least one of the subjects. Although T-cell reactive regions were located throughout the molecule, in 12 of the 18 responsive individuals, major immunogenic sites were contained within the 56 amino acids of the N-terminus, and in 11 of these individuals T-cell reactive regions were only present between amino acid positions 1 and 94. In two individuals reactive sites could be mapped in the C-terminal half of the molecule, and in five subjects, epitopes were present in both N- and C-terminal regions. HLA class I and class II DR and DQ specificities were determined serologically in 16 of the 18 individuals, and no strong pattern of association between HLA type and the T-cell immunogenic region could be detected.